Membrane-Supported, Highly-Ordered Biomaterials for Bioreactors

CTS-9307518肯塔基州研究基金会艾伦巴特菲尔德 摘要 固定化酶的方法经常由于构象变化、不利的取向和其他因素而导致活性降低。本计画将探讨一种新的技术，使膜生物反应器能更专一且均匀地附着于间隔物上。 定点诱变用于修饰半胱氨酸残基的遗传密码，然后表达变体酶（枯草杆菌蛋白酶）。 在间隔化合物的一个末端的官能团将与聚合物膜中的氨基或羟基部分反应，而另一个末端将含有硫醇以在完成固定时被氧化成二硫化物。 固定化酶和中间体结构将通过NMR、X射线荧光、自旋标记研究和其他方法来表征。 在包括水溶剂和有机溶剂的各种反应条件下考察催化行为。 酶的固定化有利于连续操作模式，并有助于生物产品的纯化。 这项基础研究可以帮助改善许多生物技术公司的生产流程。

CTS-9307518 Kentucky Research Foundation D. Allen Butterfield ABSTRACT Methods for the immobilization of enzymes often lead to reduced activity due to conformational changes, unfavorable orientations and other factors. A new technique for more specific and uniform attachment with spacers will be explored in this project on membrane bioreactors. Site-directed mutagenesis is to be used in modifying the genetic code for a cysteine residue and then expressing the variant enzyme (subtilsin). Functional groups at one terminus of the spacer compound will be reacted with amino or hydroxyl moieties in a polymeric membrane while the other terminus will contain a thiol to be oxidized to the disulfide in completing the immobilization. The immobilized enzyme and intermediate structures will be characterized by NMR, x-ray fluorescence, spin-label studies, and other methods. Catalytic behavior is to be examined under various reaction conditions including aqueous and organic solvents. Immobilization of enzymes facilitates a continuous mode of operation and aids the purification of bioproducts. This fundamental research could help improve the production processes in many biotechnology companies.